title: Peroxynitrite-induced apoptosis in epithelial (T84) and macrophage (RAW 264.7) cell lines: effect of legume-derived polyphenols (phytolens).
abstract: Peroxynitrite (ONOO-) has been proposed as a mediator of gut inflammation and as an inducer of cell death by apoptosis. Phytolens (PHY), a water-soluble extract of polyphenolic antioxidants from nonsoy legumes (Biotics Research Corp, patent pending), was evaluated as a cytoprotective agent in human colonic (T84) and murine macrophage (RAW 264.7) cell lines. In the antioxidant testing, PHY showed a significant free radical scavenging ability against 1,1-diphenyl-2-picrylhydrazyl radical (DPPH) and superoxide (O2.) radicals with an IC50 of 4.44 and 5.87 microg/ml against DPPH and O2., respectively. Apoptosis (DNA fragmentation) was measured by an ELISA technique. Cells were exposed to oxidative stress by treating them with peroxynitrite (100-300 microM) for 4 h in the presence and absence of PHY. Peroxynitrite elicited a dose-dependent increase in DNA fragmentation in both cell lines compared to the control group receiving decomposed ONOO-. PHY (10, 30, or 50 microg/ml) significantly attenuated the degree of apoptosis in T84 cells induced by ONOO- (P < 0.05). PHY (10-100 microg/ml) did not directly affect T84 cell viability or induce apoptosis after 4 h or overnight exposure. RAW 264.7 cells exposed to PHY alone (>30 microg/ml) for 4 h displayed decreased cell viability (P < 0.05) and increased apoptosis (P < 0.05). Phytolens may have beneficial effects on inflammation by attenuating peroxynitrite-induced apoptosis. The sparing of epithelial cells while compromising the viability of macrophages suggests that PHY may be beneficial in autoimmune disorders.
authors.affiliation: Department of Pediatrics and Stanley S. Scott Cancer Center, Louisiana State University Medical Center, New Orleans, USA.
authors.forename: M
authors.initials: M
authors.lastname: Sandoval
authors.forename: R A
authors.initials: RA
authors.lastname: Ronzio
authors.forename: D N
authors.initials: DN
authors.lastname: Muanza
authors.forename: D A
authors.initials: DA
authors.lastname: Clark
authors.forename: M J
authors.initials: MJ
authors.lastname: Miller
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects*; D015764:Bepridil / Q000031:analogs & derivatives / Q000378:metabolism; D001713:Biphenyl Compounds; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D003110:Colonic Neoplasms / Q000378:metabolism; D053938:DNA Fragmentation / Q000187:drug effects; D007887:Fabaceae / Q000737:chemistry; D005419:Flavonoids; D016166:Free Radical Scavengers / Q000378:metabolism; D006801:Humans; D007249:Inflammation / Q000188:drug therapy; D008264:Macrophages / Q000378:metabolism; D009566:Nitrates / Q000494:pharmacology*; D010636:Phenols / Q000494:pharmacology*; D010851:Picrates; D010946:Plants, Medicinal; D011108:Polymers / Q000494:pharmacology*; D059808:Polyphenols; D051381:Rats; D013481:Superoxides / Q000378:metabolism
chemical_list: D000975:Antioxidants; D001713:Biphenyl Compounds; D005419:Flavonoids; D016166:Free Radical Scavengers; D009566:Nitrates; D010636:Phenols; D010851:Picrates; D011108:Polymers; D059808:Polyphenols; D013481:Superoxides; C039642:peroxynitric acid; D015764:Bepridil; C004931:1,1-diphenyl-2-picrylhydrazyl